Abstract
Purpose:
The goal of this study was to interface new technologies made available through Raman spectroscopy technique with the cell culture cytotoxicity assay with the intent to develop novel noninvasive methods based on Raman spectroscopy to determine tumor viability in vivo. Raman spectroscopy is a new technology that uses monochromatic light to map diseases in a non-invasive way and can be applied to a wide variety of sample morphologies. The quantitative nature of Raman spectra, combined with the ability to provide unique fingerprints of the biochemicals present in tissue without surgical biopsy, provides objective diagnostic information for cancerous tissue analysis. We expect that Raman spectroscopy could be helpful in diagnosis and management of numerous retinal and optic nerve head conditions.
Methods:
In this study sensitivity of retinoblastoma cell lines (Y79, WERI-Rb1) to chemotherapy were compared using standard MTT assay and Raman spectroscopy technique with respect to drug ( Etoposide, Carboplatin, Vincristine) concentration and time of treatment. Raman spectra were recorded using Renishaw inVia Raman microscope with 785 nm excitation wavelength within spectral region 600-1800 cm-1.The acquired raw spectra were corrected for background influence and data were analyzed using SPSS statistical software.
Results:
Results of cytotoxicity assay indicated that all model agents acted in the mitochondrial activity by significant reducing cellular viability in the Y79 and WERI-Rb1 cells. Raman analysis results indicate clear possibility of detecting specific changes which correlated with cell survival during the treatment. We have observed significant changes over in 20 peaks and peak ratios. Moreover, following treatment, we have observed intense Raman peak within 957-959cm-1 matching the characteristic absorption peak of Hydroxyapatite crystal deposition as a marker for ocular calcifications.
Conclusions:
These preliminary results demonstrate that Raman spectroscopy has the potential to be applied in vivo to accurately detect viability of intraocular retinoblastoma lesions. It was found that there was a correlation of the Raman spectrum with the cell survival rate. These results also will provide background into the chemical/morphological makeup of the retinoblastoma tissue and will be used to develop diagnostic algorithms for portable Raman clinical diagnostic tool.
Keywords: 703 retinoblastoma •
449 cell survival •
552 imaging methods (CT, FA, ICG, MRI, OCT, RTA, SLO, ultrasound)